Experimental infection and protection against ... - TI Pharma

136 Chapter 7
NF135.C10 NF54
Re-started cultures until CHMI 7 306
Period 2009-2010
Infection 74% (62-87) 86% (78-94)
Oocysts 12 (7.3-16) 27 (22-33)
Sporozoites/mosquito x10 3 39 (18-60) 99 (74-124)
CHMI (April 2010)
Infection 100% 100%
Oocysts 5.6 17
Sporozoites/mosquito x10 3 12.5 69
Gametocyte male: female ratio 1:5 1:3
Drug sensitivity (IC50)
dihydroartemisinin 3.4 nM 9.9 nM
lumefantrine 89 nM 78 nM
proguanil 21 µM 27 µM
atovaquone 0.3 nM 0.6 nM
chloroquine 201 nM 24 nM
Table 1. Mosquito infection and drug sensitivity profile of NF135.C10 and NF54 in
the period 2009-2010 and for the specific batches used in this CHMI. Data are
displayed for the period 2009-2010 (mean (95% CI)) after 26 and 39 standard
dissections respectively, from 10 mosquitoes per dissection. Half-inhibitory
concentrations (IC50) are means from three independent experiments.
17H12, eBioscience )) or 2) IFNγ FITC, in permeabilization Medium B (Caltag).
Cells were read on a CyAn ADP 9-color flow cytometer (Beckman-Coulter) and
analysed using FlowJo software (Tree Star, Inc.) version 9.2. Gating of cytokinepositive
cells was performed based on the Median Fluorescent Intensity (MFI) of
cytokine negative PBMCs for each volunteer, time point and stimulus.
Statistical analysis
Data analysis was performed using GraphPad Prism5 software. Differences in
parasite kinetics between subjects in the NF135.C10 and NF54 group were
analysed using the non-parametric Mann-Whitney test. A two-sided P value of
less than 0.05 was considered statistically significant.
Results
Generation and characterization of NF135.C10
Clinical isolates of asexual Pf parasites were obtained by culturing blood from Pfmalaria
patients from hospitals in The Netherlands. We adapted 74 different